APPROFONDIMENTO DELLE CONOSCENZE SULL'ECOLOGIA DEI DITTERI CHIRONOMIDI DELLE ACQUE INTERNE ITALIANE

The general aim of this work was to study lakes\u2019s benthic communities, particularly Chironomids (Diptera Chironomidae), to select the environmental factors correlated to their diversity and distribution and to provide information about their ecology. A self-organizing map analysis (SOM) was carried out to ordinate sites (samples from 50 Italian lakes) according to macroinvertebrate community composition. Environmental variables were also included in the two dimension maps to highlight which factors are associated with site-taxa distribution (dissolved oxygen and nutrient content were selected). Within species with an indicator role of hypoxia, high nutrient amount, inorganic and organic pollution, the Chironomid Chironomus riparius Meigen 1804 was selected. A first series of experiments were carried out in lab on IV-instar larvae to investigate the role of the respiratory pigment hemoglobin and of the enzyme alcohol dehydrogenase in survive hypoxia and anoxia. A second series of experiments were carried out always on IV-instar larvae to investigate the role of five candidate genes (hsp10, hsp40, hsp70, hsc70 and cyp450) in toxic resistance (Cu and azadirachtin) (Real Time PCR analysis). Overall more information on autoecology of Chironomids was given by this study, and new data on potential biomarkers useful in biomonitoring of lakes

ALCOHOL HARM-REDUCTION APPROACHES ACROSS DIFFERENT POPULATIONS

Context. Certain populations, such as socially marginalized individuals and late adolescents (i.e., aged 17-19), are more prone than others to experience severe alcohol- related harm. However, these populations are often not interested in attaining alcohol abstinence or reducing alcohol consumption to moderate levels. Thus, their needs are not met within traditional primarily abstinence-based service options. In response, harm- reduction approaches have begun to be applied with these populations. The overall aim of this thesis was to contribute to the research on alcohol-related harm reduction in these populations by conducting 5 distinct studies. The two first studies were conducted with chronically homeless individuals with alcohol problems in the US: Study 1 examined their perceptions of twelve-step mutual-help groups (TMGs) and their association with treatment attendance, motivation and alcohol outcomes, whereas study 2 examined this population's use of safer-drinking strategies. Study 3 evaluated substance-use and related problems following exposure to a harm-reduction drop-in center that allows alcohol consumption onsite among socially marginalized alcohol and other drug (AOD) users in Switzerland. Studies 4 and 5 were conducted among US and Swedish high school seniors and prospectively tested a) the moderating effect of drinking intentions on the association between use of protective behavioral strategies (PBS) and alcohol outcomes, and b) the moderating effect of PBS use on the link between alcohol expectancies and alcohol outcomes. Findings. Study 1 found that most chronically homeless individuals with alcohol problems endorsed negative perceptions of TMGs, which were in turn associated with less treatment attendance. Study 2 found that chronically homeless individuals with alcohol dependence were interested in drinking safer and were able to commit to using safer-drinking strategies. Study 3 showed that exposure to a harm-reduction drop-in center that allows alcohol consumption onsite was followed by significant decrease in alcohol use and related problems among socially marginalized AOD users in Switzerland, with greater attendance being related to additional improvement in mental health-related quality of life (QoL) and decrease in drug-related problem severity. Finally, studies 4 and 5 showed that use of PBS was related to future decreases in alcohol use and related consequences among US and Swedish high school seniors. Findings also showed significant interactions between drinking intentions and PBS use (study 4), and between alcohol expectancies and PBS use (study 5). Conclusion. Taken together, findings confirmed that socially marginalized individuals and high school seniors show low interest in traditional approaches aiming abstinence or use reduction. Findings documented, however, that these distinct populations embrace alcohol harm-reduction approaches and that these approaches appear to be related to decreases in alcohol-related harm. Taken together, these findings suggest that alcohol harm-reduction approaches are promising ways to decrease alcohol- related harm across different populations and different cultures (i.e., US, Sweden, Switzerland). -- Contexte. Malgré leurs besoins importants en matière de traitement alcoologique, les personnes socialement marginalisées sont souvent sous-traitées par les traitements visant l'abstinence ou la consommation modérée. Similairement, les approches préventives promouvant l'abstinence ne suscitent généralement que peu d'intérêts et d'effets positifs au sein des jeunes (gymnasiens par exemple), qui sont pourtant connus pour leur usage d'alcool à risque. En réponse, les interventions ciblant ces populations intègrent de plus en plus des principes de réduction des risques et des méfaits. L'objectif général de ce travail de thèse était de contribuer à la recherche dans ce domaine en réalisant 5 études distinctes. Les deux premières études ont été conduites au sein de personnes sans-abris chroniques présentant un problème d'alcool aux États-Unis: L'étude 1 a examiné leurs perceptions des groupes d'entraide mutuelle en douze étapes (GEMs), et l'association entre ces perceptions et la fréquentation des traitements, la motivation et l'usage d'alcool. L'étude 2 a examiné l'usage des stratégies de consommation plus sûre au sein de cette population. L'étude 3 a évalué l'évolution de l'usage de substances et des problèmes associés après l'exposition à un espace d'accueil de jour tolérant la consommation d'alcool au sein de personnes socialement marginalisées en Suisse. Enfin, les études 4 et 5 ont été conduites parmi des gymnasiens aux États-Unis et en Suède et ont testé prospectivement a) l'effet modérateur des intentions de consommer sur le lien entre l'utilisation de stratégies de protection comportementales (SPCs) et l'usage d'alcool, et b) l'effet modérateur de l'usage de SPCs sur le lien entre les attentes liées à l'usage d'alcool et la consommation d'alcool. Résultats. L'étude 1 a montré que la plupart des participants avaient des perceptions négatives des GEMs et que ces perceptions étaient associées négativement à la fréquentation des traitements. L'étude 2 a montré que les participants étaient intéressés à réduire les risques liés à leur usage d'alcool et qu'ils pouvaient s'engager à utiliser des stratégies de consommation plus sûre. L'étude 3 a montré que les participants ont diminué leur usage d'alcool et les problèmes associés au cours de l'évaluation et que chaque mois supplémentaire passé au sein de l'espace d'accueil était associé à une diminution supplémentaire de la sévérité des problèmes liés aux drogues et à une amélioration de la qualité de vie liée à la santé mentale. Les études 4 et 5 ont indiqué que l'usage de SPCs était prospectivement associé à une diminution de la consommation d'alcool et des conséquences associées. Les résultats ont également montré des interactions significatives entre l'usage des SPCs et a) les intentions de consommer (étude 4), et b) les attentes liées à l'alcool (étude 5). Conclusion. Globalement, les études comprises dans ce travail de thèse confirment que les approches visant l'abstinence et la consommation modérée ne suscitent que peu d'intérêt au sein des personnes socialement marginalisées et des gymnasiens. Notre travail indique toutefois que ces populations acceptent les approches visant la réduction des risques et des méfaits et que celles-ci semblent être associées à une diminution des méfaits liés à l'usage d'alcool. De manière générale, ces résultats suggèrent que ces approches sont prometteuses au sein de différentes populations issues de cultures diverses (États-Unis, Suède, Suisse)

Modulating the WNT pathway in Drosophila models of Cornelia de Lange Syndrome

Cornelia de Lange syndrome (CdLS) is a rare genetic disorder affecting neurodevelopment and the gastrointestinal and musculoskeletal systems. CdLS is caused by mutations within NIPBL, SMC1A, SMC3, RAD21, HDAC8 and BRD4 genes. These genes codify for the cohesin complex (or associated proteins), a multiprotein structure playing a role in chromatid adhesion, DNA repair and gene expression regulation. Our laboratory has shown that a strong correlation exists between cohesin complex function and WNT signalling, an intracellular pathway involved in regulation of expression of several genes controlling cell division and embryonic development. Recently, it has been observed that in nipblb- and smc1a-loss-of-function zebrafish embryos, in NIPBL- and SMC1A- mutated patient fibroblasts and in CdLS murine neural stem cells (NSCs) similar patterns of canonical WNT pathway alterations and cyclinD1 downregulation are present. Indeed, zebrafish embryos adverse phenotype was rescued by chemical activation of the WNT pathway. Drosophila melanogaster is an inexpensive model to study CdLS and to screen in vivo for therapeutic compounds. Therefore, we have used flies\u2019 strains mutated in nipped-B and hdac3 genes (respectively NIPBL and HDAC8 in humans) for assessing the existing correlation between cohesin complex and WNT pathway. Moreover, we have selected D. melanogaster mutants to screen for chemicals that revert the CdLS associated-phenotypes efficiently. In particular, we have tested WNT activators in order to investigate which effects they have on the mutated flies, assessing body weight and changes in brain structures (i.e. mushroom bodies) and possibly select compounds to test on vertebrate models. Funding: This work has been supported by Fondazione Cariplo, grant 2015-0783 to Valentina Massa

Circular dichroism and bilayer splitting in the normal state of underdoped (Pb,Bi)2_2Sr2_2(Cax_xY1−x_{1-x})Cu2_2O8+δ_{8+\delta} and overdoped (Pb,Bi)2_2Sr2_2CaCu2_2O8+δ_{8+\delta}

We report an ARPES investigation of the circular dichroism in the first Brillouin zone (BZ) of under- and overdoped Pb-Bi2212 samples. We show that the dichroism has opposite signs for bonding and antibonding components of the bilayer-split CuO-band and is antisymmetric with respect to reflections in both mirror planes parallel to the c-axis. Using this property of the energy and momentum intensity distributions we prove the existence of the bilayer splitting in the normal state of the underdoped compound and compare its value with the splitting in overdoped sample. In agreement with previous studies the magnitude of the interlayer coupling does not depend significantly on doping. We also discuss possible origins of the observed dichroism.Comment: 4 RevTex pages, 4 EPS figure

Nutritional management of search and rescue dogs

Dogs used for search and rescue (SAR) may experience continuous micro-traumas that predispose them to skeletal disorders. The aim of the present study was to evaluate the effect of diet on osteo-articular apparatus in healthy SAR dogs. A total of sixteen SAR dogs were divided into two groups (low supplementation (LS) and high supplementation (HS)) and were fed for 3 months with two experimental diets, characterised by the same protein and energy density, but different inn-3 PUFA (6·2v. 8·4 % of metabolisable energy), chondroitin sulfate (219·8v. 989·0 mg/kg DM) and glucosamine (769·2v. 1318·7 mg/kg DM) in the LS and HS groups, respectively. At recruitment all dogs showed no joint inflammation signs, except four that showed mild symptoms. Haematology and serum biochemistry were performed every 30 d. Joint status was scored by physical and lameness evaluations. The sampling effect analysis showed potential beneficial effects by a decrease in a specific marker of membrane integrity (creatine kinase; CK). Comparing groups, glucose was significantly higher and CK was significantly lower in the HS group; however, in both cases the levels of these parameters fell in the normal range. At the end of the experiment, erythrocytes, Hb and packed cell volume were significantly higher in the HS compared with the LS group. These could result in an improvement in dogs' performance, even if this aspect was not investigated in the present study. Concerning joint evaluation (pain on manipulation, lameness and range of motion), no statistically significant differences were detected between the groups and within the experimental period

Influence of Pasture on Stearoyl-CoA Desaturase and miRNA 103 Expression in Goat Milk: Preliminary Results

The effect of pasture on the stearoyl-CoA desaturase (SCD) and miRNA 103 expression was evaluated on dairy goats divided into two homogeneous groups (G, grazing, and S, stable). Group S was housed in a stall and received alfalfa hay as forage, while group G was led to pasture. The goats of both the groups received the same amount of concentrate. Milk yield did not di↵er statistically between the groups. Group G showed significantly higher fat (4.10% vs. 2.94%, p < 0.01) and protein percentage (3.43% vs. 3.25%; p < 0.05) than group S. Among milk fatty acids, group S showed significantly higher levels of saturated fatty acids (SFA) and lower values of mono-unsaturated fatty acid (MUFA). The percentages of polyunsaturated fatty acid (PUFA) were not different between groups even if pasture significantly affected the percentages of C18:3 and total omega 3. In group G, total CLAs were twice than in group S (0.646% vs. 0.311%; p < 0.01) mainly due to the differences in CLA cis9 trans 11 (0.623% vs. 0.304%; p < 0.01). Milk total CLA in grazing group was significantly (p < 0.01) higher in August according to the highest value of both linoleic and alfa-linolenic acids in the pasture. In grazing animals, SCD expression decreased from April to June, increased in July and decreased again in August, while it was almost unvaried along the trial in group S. By contrast, the expression of miRNA 103 showed a similar trend for both groups, decreasing from April to June, increasing in July and falling down in August. To our knowledge, this is the first observation of the effects of pasture on miRNA expression in milk from ruminant species

The ground and ionic states of cyclohepta-1,3,5-triene and their relationship to norcaradiene states. New 1H and 13C NMR spectra, and analysis of a new experimental photoelectron spectrum by ab initio methods

The strong inter-relationship between cyclohepta-1,3,5-triene (CHT) and norcaradiene (NCD) observed in some reactions, has been extended to include the energy surfaces for some low-lying ionic states. Whilst equilibrium structures of 2A/ symmetry containing the CHT skeleton were routinely found, the structures emerging with 2A// symmetry were found to be NCD ionic states. Surface studies, by variation of the C1 to C6 distance, showed minima for both state symmetries. Curve crossing which occurs in CS symmetry, is avoided by distortion to C1 symmetry. The CHT → NCD structural change is attributed to initial conrotatory closure of the singly occupied molecular orbital. A new synchrotron-based study of the photoelectron spectrum (PES) for CHT up to 25 eV shows little vibrational structure. We have assigned the overall PES up to 17 eV in considerable detail, using a variety of theoretical methods. The vertical ionization energy (VIE) sequence in the PES for CHT ions, is predicted to be: 12A/ < 12A// < 22A/ < 22A//. The calculated lowest ionic state, with lowest vibrational frequency 87 cm-1, leads to a high density of vibrational states. The Franck-Condon (FC) envelopes of the two lowest PES bands have been analysed. The identity of the PES spectrum, as derived from CHT rather than NCD, was demonstrated by 1H and 13C nuclear magnetic resonance (NMR) spectra of the sample; this agrees with the predicted PES spectra of CHT and NCD. The NCD predicted spectrum shows significant differences from CHT.PostprintPostprintPeer reviewe

Drosophila melanogaster as a model to study WNT pathway alteration in Cornelia de Lange Syndrome

Introduction: Cornelia de Lange syndrome (CdLS) is a rare genetic disorder affecting neurodevelopment, gastrointestinal and musculoskeletal systems. CdLS is caused by mutations within NIPBL, SMC1A, SMC3, RAD21, or HDAC8 genes. These genes codify for the cohesin complex, a multiprotein structure playing a role in chromatid adhesion, DNA repair and gene expression regulation. It has been demonstrated that a strong correlation exists between cohesin complex function and WNT signalling. Recently, it has been observed that chemical activation of the WNT pathway in nipblb-loss-of-function zebrafish embryos and in NIPBL-mutated patient fibroblasts rescued the adverse phenotype. Both embryos and fibroblasts present similar patterns of canonical WNT pathway alterations and CCND1 downregulation. Materials and Methods: Drosophila melanogaster is an inexpensive model to study CdLS and to screen in vivo for therapeutic compounds. Therefore, we have selected fly strains mutated in nipped-B and hdac3 genes (respectively NIPBL and HDAC8 in humans) for assessing the existing correlation between cohesin complex and WNT pathway and to screen for chemicals that revert the CdLS associated-phenotypes efficiently. Results: We have confirmed that mutated flies weight 5% less than wild type. Moreover, we have tested lithium chloride (LiCl) as WNT activator, demonstrating that 250mM is the highest concentration tolerated. Conclusions: Hence, we hypothesize that WNT pathway activation could improve mutant phenotype. We will be testing different doses of LiCl and other WNT activator to assess whether some of those chemical compounds could revert the syndrome-associated phenotype